Automated High-Throughput Seed Sampler And Methods Of Sampling, Testing And Bulking Seeds

ABSTRACT

An automated method for operating an automated seed sampling system having a seed loading station, a seed transport subsystem, and a seed sampling station generally includes sensing whether individual seeds are successfully isolated from a bulk of seeds at the seed loading station, and sensing whether the isolated seeds are properly positioned by the seed transport subsystem adjacent the seed sampling station in preparation for removing tissue from the isolated seeds. In some aspects, the method further includes analyzing the tissue removed from the seeds for presence or absence of at least one characteristic, and selecting seeds based on presence or absence of the at least one characteristic.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.15/411,531, filed Jan. 20, 2017, which is a continuation of U.S. patentapplication Ser. No. 14/635,738 (U.S. Pat. No. 9,551,636), filed Mar. 2,2015, which is a continuation of U.S. patent application Ser. No.13/897,024 (U.S. Pat. No. 8,997,398), filed May 17, 2013, which is acontinuation of U.S. patent application Ser. No. 13/556,742 (U.S. Pat.No. 8,443,545), filed Jul. 24, 2012, which is a continuation of U.S.patent application Ser. No. 13/251,993 (U.S. Pat. No. 8,245,439), filedOct. 3, 2011, which is a continuation of U.S. patent application Ser.No. 12/128,279 (U.S. Pat. No. 8,028,469), filed May 28, 2008. U.S.patent application Ser. No. 12/128,279 claims the benefit of U.S.Provisional Application Ser. No. 60/940,788, filed May 30, 2007, and isalso a continuation-in-part of U.S. patent application Ser. No.11/680,180 (U.S. Pat. No. 7,998,669), filed Feb. 28, 2007, which claimsthe benefit of U.S. Provisional Application Ser. No. 60/778,830, filedMar. 2, 2006. The entire disclosures of the above applications areincorporated herein by reference.

FIELD

This disclosure generally relates to systems and methods for takingsamples from biological materials such as seeds.

BACKGROUND

The statements in this section merely provide background informationrelated to the present disclosure and may not constitute prior art.

In plant development and improvement, genetic improvements are made inthe plant, either through selective breeding or genetic manipulation,and when a desirable improvement is achieved, a commercial quantity isdeveloped by planting and harvesting seeds over several generations. Notall seeds express the desired traits and, thus, these seeds need to beculled from the population. To hasten the process of bulking up thepopulation, statistical samples are taken and tested to cull seeds fromthe population that do not adequately express the desired trait.

U.S. patent application Ser. No. 11/680,180 (filed Feb. 28, 2007); U.S.patent application Ser. No. 11/213,430 (filed Aug. 26, 2005); U.S.patent application Ser. No. 11/213,431 (filed Aug. 26, 2005); U.S.patent application Ser. No. 11/213,432 (filed Aug. 26, 2005); U.S.patent application Ser. No. 11/213,434 (filed Aug. 26, 2005); U.S.patent application Ser. No. 11/213,435 (filed Aug. 26, 2005); U.S.patent application Ser. No. 11/680,180 (filed Feb. 27, 2007); and U.S.patent application Ser. No. 11/680,611 (filed Feb. 27, 2007), which areincorporated herein by reference in their entirety, disclose apparatusand systems for the automated sampling of seeds as well as methods ofsampling, testing and bulking seeds.

SUMMARY

The present disclosure relates to systems and methods of separatingseeds from a plurality of seeds, extracting a sample from each seed,sorting the extracted samples and corresponding seeds respectively towells in sample trays and seed trays, and mapping the respective wellsto track each sample with the seed from which it was extracted. Themethods are particularly adapted for automation, which permits a greatersampling and sorting efficiency and throughput rate than was previouslypractical.

In various embodiments, the present disclosure provides an automatedsystem for sampling and sorting at least one seed from a plurality ofseeds. The system includes a seed loading station for separating atleast one seed from a plurality of seeds in a bulk seed hopper, animaging station for collecting image data of the at least one seed, anda seed orientation station for independently positioning and retainingeach seed in a desired orientation based on the collected image data.The system also includes a seed sample and sort station for extracting atissue sample from each seed, sorting each tissue sample to a sampletray and sorting each sampled seed to a seed tray.

In various other embodiments, the present disclosure provides anautomated, high-throughput method for extracting sample material fortesting from individual seeds in a population of seeds. The methodincludes separating at least one seed from a plurality of seeds in apopulation, collecting image data for the at least one seed,independently positioning the at least one seed in a desired orientationbased on the collected image data, extracting a tissue sample from theat least one seed, and sorting the tissue sample to a sample tray andsorting the sampled seed to a seed tray.

In yet other various embodiments, the present disclosure provides anautomated system for sequentially removing sample material fromindividual ones of a plurality of seeds while preserving the germinationviability of the seeds. The system includes a seed loading station forseparating and retaining sets of seeds from a plurality of seeds in abulk seed hopper, an imaging station for collecting image data of theretained sets of seeds, and a seed orientation station for independentlypositioning each seed in each seed set in a desired orientation based onthe collected image data. The system also includes a seed sample andsort station for extracting a tissue sample from each seed in each seedset, sorting each tissue sample to a sample tray and sorting eachsampled seed to a seed tray.

In still yet other various embodiments, the present disclosure providesan automated, high-throughput method for sequentially extracting samplematerial for testing from a plurality of seeds while preserving thegermination viability of the seeds. The method includes separating setsof seeds from a plurality of seeds in a bulk seed hopper. Each set ofseeds is then presented for retention by a respective one of a pluralityof rotary vacuum cup banks at a seed loading station of a seed samplingand sorting system. Each rotary vacuum cup bank includes a plurality ofrotary vacuum cup devices. The method additionally includes collectingimage data of each set of seeds retained by each rotary vacuum cup bankat an imaging station of the seed sampling and sorting system. Themethod further includes independently positioning each seed in the setin a desired orientation based on the collected image data at a seedorientation station of the seed sampling and sorting system. The methodstill further includes extracting a sample from each seed in each set ofseeds; and sorting each sample to a sample tray and sorting each sampledseed to a seed tray at a seed sample and sort station of the seedsampling and sorting system.

In still other various embodiments, the present disclosure provides aseed loading station of an automated seed sampling and sorting system.The seed loading station includes a separating wheel for separatingseeds from a plurality of seeds in a bulk hopper. Additionally, the seedloading station includes a tube shuttle having a plurality of firsttransfer tubes extending from a plurality of openings in the tubeshuttle. The tube shuttle is structured and operable to incrementallypositioning each of the first transfer tubes under the separating wheelsuch that each of the first transfer tubes receives a seed from theseparating wheel.

In other various embodiments, the present disclosure provides a seedsample and sort station of an automated seed sampling and sortingsystem. The seed sample and sort station includes a press plate bankincluding a number of press plates equal to a number of seed retentiondevices of an automated seed sampling and sorting system. Each seedretention device retains a respective seed. The seed sample and sortstation additionally includes a linear actuator to which the press platebank is mounted. The linear actuator is controllable to lower the pressplate bank such that each press plate engages a friction plate of acorresponding one of the retention devices and moves the respectiveseeds downward to a sampling location. The seed sample and sort stationfurther includes a plurality of independently controlled grip and chipassemblies. Each grip and chip assembly includes a seed grippingmechanism for firmly holding a respective one of the seeds at therespective sampling location as a sample is extracted from therespective seed. Each grip and chip assembly additionally includes asample extraction mechanism for extracting the sample from eachrespective seed.

In still other embodiments, methods are provided for removing tissuefrom multiple individual seeds. In one example embodiment, a method forremoving tissue from multiple individual seeds generally includesloading multiple individual seeds in a seed transport, orienting themultiple individual seeds in the seed transport substantiallysimultaneously, and removing tissue from the oriented multipleindividual seeds.

In another example embodiment, an automated method for sampling seedsincludes separating individual seeds from a plurality of seeds, imagingthe separated seeds, and removing tissue samples from the imaged seeds.

In other embodiments, seed sampling systems are provided. In one exampleembodiment, a seed sampling system generally includes a seed transportconfigured to hold multiple individual seeds together as a group andtransport the multiple individual seeds together as the group, and toallow the multiple individual seeds to be oriented while the multipleindividual seeds are being held in the seed transport; and a seedsampling subsystem configured to remove tissue from the orientedmultiple individual seeds.

In another example embodiment, an automated system for sampling seedsincludes a seed loading station for separating individual seeds from aplurality of seeds held in a seed hopper, an imaging station configuredto receive the separated seeds from the seed loading station and collectimage data of the received seeds, and a seed sampling subsystemconfigured to remove tissue samples from the seeds after the image dataof the seeds is collected.

Further areas of applicability of the present teachings will becomeapparent from the description provided herein. It should be understoodthat the description and specific examples are intended for purposes ofillustration only and are not intended to limit the scope of the presentteachings.

DRAWINGS

The drawings described herein are for illustration purposes only and arenot intended to limit the scope of the present teachings in any way.

FIG. 1 is an isometric view of a seed sampling system in accordance withvarious embodiments of the present disclosure.

FIG. 2 is a block diagram of a top view of the seed sampling systemshown in FIG. 1.

FIG. 3A is an isometric view of a seed loading station (absent systemsupport structure) of the seed sorter system shown in FIG. 1, inaccordance with various embodiments of the present disclosure.

FIG. 3B is a schematic of a queuing stack of the seed loading stationshown in FIG. 3A, in accordance with various embodiments of the presentdisclosure.

FIG. 3C is a sectional side view of an elevator hopper of the seedloading station shown in FIG. 3A, in accordance with various embodimentsof the present disclosure.

FIG. 4A is an isometric view illustrating a seed transport subsystem(absent system support structure), of the seed sorter system shown inFIG. 1, including a transport carousel having one of a plurality ofrotary vacuum cup banks mounted thereto, in accordance with variousembodiments of the present disclosure.

FIG. 4B is an isometric view of a rotary vacuum cup included in therotary vacuum cup bank shown in FIG. 4A, in accordance with variousembodiments of the present disclosure.

FIG. 5A is an isometric view of an imaging station (absent systemsupport structure), of the seed sorter system shown in FIG. 1, inaccordance with various embodiments of the present disclosure.

FIG. 5B is an exemplary schematic illustrating a 360° plane in which theorientation of seed tips of a plurality of seeds are determined at theimaging station shown in FIG. 5A.

FIG. 6A is an isometric view of a seed orientation station (absentsystem support structure), of the seed sorter system shown in FIG. 1, inaccordance with various embodiments of the present disclosure.

FIG. 6B is an exemplary schematic illustrating a 360° plane in which theseeds are controllably rotated at the seed orientation station, shown inFIG. 6A, such that the seed tips of each seed have a desiredorientation.

FIG. 6C is an isometric partial view of the seed orientation stationshown in FIG. 6A, illustrating a seed purge hopper, in accordance withvarious embodiments of the present disclosure.

FIG. 7 is an isometric view of the seed sampling system, shown in FIG.1, illustrating a seed sample and sort station, in accordance withvarious embodiments of the present disclosure.

FIG. 8A is an isometric view of a seed sampling subsystem (absent systemsupport structure), of the seed sample and sort station, shown in FIG.7, in accordance with various embodiments of the present disclosure.

FIG. 8B is an isometric view of the seed grip and chip assembly of theseed sampling subsystem, shown in FIG. 8A, in accordance with variousembodiments of the present disclosure.

FIG. 8C is an isometric view of an exemplary seed gripping fingerincluded in the seed grip and chip assembly, shown in FIG. 8B, inaccordance with various embodiments of the present disclosure.

FIG. 8D is a top view of a seed gripping mechanism of the seed grip andchip assembly, shown in FIG. 8B, in accordance with various embodimentsof the present disclosure.

FIG. 8E is an isometric view of an exemplary cutting wheel of the seedgrip and chip assembly, shown in FIG. 8B, in accordance with variousembodiments of the present disclosure.

FIG. 9A is a front view of a seed and sample sorting subsystem, of theseed sample and sort station, shown in FIG. 7, in accordance withvarious embodiments of the present disclosure.

FIG. 9B is a side view of seed and sample sorting subsystem shown inFIG. 9A.

FIG. 9C is diagonal view of a sample extraction nozzle manifold of theseed and sample sorting subsystem shown in FIGS. 9A and 9B, inaccordance with various embodiments of the present disclosure.

FIG. 9D is an isometric view of a sample tray platform and X-Ytranslation stage included in the seed and sample sorting subsystemshown in FIGS. 9A and 9B.

FIG. 9E is an isometric view of a seed tray platform and X-Y translationstage included in the seed and sample sorting subsystem shown in FIGS.9A and 9B.

DETAILED DESCRIPTION

The following description is merely exemplary in nature and is in no wayintended to limit the present teachings, application, or uses.Throughout this specification, like reference numerals will be used torefer to like elements.

FIGS. 1 and 2 illustrate an automated seed sampling system 10, inaccordance with various embodiments of the present disclosure. The seedsorter system 10 includes a seed loading station 100, a seed transportsubsystem 200, a seed imagining station 300, a seed orientation station400, a seed sampling and sort station 500 and a central controllersystem (CCS) 700 that controls the operation of the seed sorter system10.

Generally, the seed sampling system 10 is structured and operable torepetitiously separate a select number of seeds, e.g., sets of eightseeds at a time, from a bulk of seeds within a bulk seed hopper 104(e.g., FIG. 3A, etc.) at the seed loading station 100. Additionally, theseed sampling system 10 is structured and operable to image each set ofseeds at the imaging station 300. The images collected at the imagingstation 300 can be any desirable type of images. For example, the imagescan be visual images, near infra-red (NIR) images or NMR/MRI images, orany other type images. In various embodiments, the imaging station 300collects at least one digital image of each set of seeds. The image datacollected of each set of seeds is communicated to a CCS 700 where theimage data is analyzed to determine the orientation, e.g., ‘tip out’ or‘crown out’. The seed sampling system 10 is further structured andoperable to orient each set of seeds in a desired orientation, based onthe images of each respective set of seeds, at the orientation station400. The seed sampling system 10 is still further structured andoperable to extract a sample (e.g., a tissue sample, etc.) from aselected area, e.g., the crown, of each seed in each set of seeds.Further yet, the seed sampling system 10 is structured and operable tothen collect the extracted samples in a plurality of sample trays 14 andsort the respective sampled seeds into a plurality of seed trays 18 atthe seed sample and sort station 500.

Once a set of seeds is separated from the bulk of seeds at the seedloading stations, as described below, the seeds are loaded onto one of aplurality of rotary vacuum cup (RVC) banks 204. The respective set ofseeds is then sequentially positioned adjacent each of the imagingstation 300, the orientation station 400 and the sample and sort station500, via the seed transport subsystem 200. More specifically, the seedtransport subsystem 200 includes an automated transport carousel 208 towhich the plurality of the RVC banks 204 are mounted. The automatedtransport carousel 208 (FIG. 4A) is driven by a motor (not shown), e.g.,a stepper motor, that incrementally rotates the transport carousel 208to sequentially advance each RVC bank 204 to each of the stations 100,300, 400 and 500. Therefore, each set of seeds is retained by arespective RVC bank 204 and transported to positions adjacent each ofthe imaging station 300, the orientation station 400 and the sample andsort station 500 by the incremental rotation of the transport carousel208.

The operation of the seed sorter system 10 is generally completelycontrolled and automated by the CCS 700 such that the operationsperformed by the imaging station 300, the orientation station 400 andthe sample and sort station 500 occur substantially without need forhuman interaction, intervention or control. However, such actions asloading the seeds into the bulk seed hopper 104 and/or physicallymanipulating and/or changing the sample trays 14 and seed trays 18(either individually or collectively) can be performed manually withhuman participation.

Generally, the CCS 700 includes one or more processors and/ormicroprocessors, and one or more electronic data storage devicesutilized to store and execute various custom programs, applicationsand/or algorithms to effectuate the operation of the seed sorter system10. Accordingly, the CCS 700 can comprise a specially programmedcomputer, or computer system, in communication with associated systemdevices (not shown) that enable communication with and control over theoperations of the various stations 100, 300, 400 and 500 and thetransport subsystem 200 of the seed sorter system 10. Although the CCS700 is exemplarily illustrated in FIG. 2 as a single unit, the CCS 700can be a single computer based system or a plurality of computer basedsubsystems networked together to coordinate the simultaneous operationsof the seed sorter system 10, as described herein. For example, invarious embodiments, the CCS 700 can include a main controller subsystemnetworked together with a plurality of peripheral controller subsystems(not shown), e.g., a peripheral controller subsystem for each station100, 300, 400, 500 and transport subsystem 200. Each peripheralcontroller subsystem can include one or more processors, microprocessorsand electronic data storage devices that effectuate communication withvarious seed sorter system components, e.g., sensors, devices,mechanisms, motors, tools, etc., and together with the main controllersubsystem cooperatively operate all the stations, systems and subsystemsof the seed sampler system 10. Or alternatively, the CCS 700 cancomprise a single computer communicatively connected to all the varioussystem components to cooperatively operate all the stations, systems andsubsystems of the seed sampler system 10.

As described above, the CCS 700 communicates with various seed sortersystem components that include various system sensors. The systemsensors operate to detect conditions of interest during operation of theseed sorter system 10 and communicate that information to the CCS 700.With this information, the CCS 700 generates control commands thateffectuate the operations and actions taken by the various stations andcomponents of the seed sorter system 10. For example, a sensed conditioncan concern: the successful isolation of sets of seeds from the seedhopper 104; the successful retention, or loading, of the each of theseeds by a respective RVC bank 204; the proper positioning of eachloaded bank of seeds adjacent each respective station 300, 400 and 500;the status (for example, position, location, vacuum, pressure, and thelike) of various component parts of the various stations 100, 300, 400and 500; operation, maintenance, performance, and error feedback fromthe various components of each station 100, 300, 400 and 500 (separatefrom, or perhaps comprising or in conjunction with, collected data); andthe like. More specifically, sensor information that is collected andprocessed for use in controlling operation of the seed sorter system 10can include such information as: device or component status; errorsignals; movement; stall; position; location; temperature; voltage;current; pressure; and the like, which can be monitored with respect tothe operation of each of the stations, subsystems and associatedcomponents of the seed sorter system 10.

It should be understood that the seed sorter system 10, as shown anddescribed herein, includes various stationary braces, beams, platforms,pedestals, stands, etc., to which various components, devices,mechanisms, systems, subsystems, assemblies and sub-assemblies describedherein are coupled, connected and/or mounted. Although such braces,beams, platforms, pedestals, stands, etc., are necessary to theconstruction of the seed sampler system 10, description of theirplacement, orientation and interconnections are not necessary for oneskilled in the art to easily and fully comprehend the structure,function and operation of the seed sampler system 10. Particularly, suchbraces, beams, platforms, pedestals, stands, etc., are clearlyillustrated throughout the figures and, as such, their placement,orientation and interconnections are easily understood by one skilled inthe art. Therefore, for simplicity, such braces, beams, platforms,pedestals, stands, etc. will be referred to herein merely as systemsupport structures, absent further description of their placement,orientation and interconnections.

Referring now to FIGS. 3A, 3B and 3C in various embodiments, the seedloading station 100 includes the seed hopper 104 and a separating wheel108. The separating wheel 108 is mounted for rotation in a verticalplane such that a portion of the separating wheel 108 extends into aninterior reservoir of the seed hopper 104. Another portion of theseparating wheel 108 extends outside of the seed hopper 104 such that aface 110 of the separating wheel 108 is positioned adjacent a seedcollector 114. The seed separating wheel 108 includes a plurality ofspaced apart recessed ports 118 that extend through the face 110 and arecommunicatively coupled to a vacuum system (not shown) such that avacuum can be provided at each of the recessed ports 118.

To initiate operation of the seed sampler system 10, seeds to be sampledand tested are placed in the seed hopper 104 interior reservoir and avacuum is provided to at least some of the recessed ports 118, e.g., therecessed ports 118 in the face 110 of the portion of the separatingwheel 108 extending into the interior reservoir of the seed hopper 104.The seed separating wheel 108 is then incrementally rotated, via anindexing motor 122, such that recessed ports 118 sequentially rotatethrough the interior reservoir of the seed hopper 104, out of the seedhopper 104, and past seed collector 114 before re-entering the interiorreservoir of the seed hopper 104. As the separating wheel incrementallyrotates and the recessed ports 118 incrementally pass through the seedhopper 104 interior reservoir, individual seeds are picked up and heldat each recessed port 118 by the vacuum provided at the respectiverecessed ports 118. As the separating wheel 108 incrementally rotates,the seeds are carried out of the seed hopper 104 to the seed collector114 where each seed is removed from the face 110 of the separating wheel108.

In various embodiments, the seed collector 114 includes a wiper (notshown) that physically dislodges each seed from the respective recessedport 118 as the separating wheel 108 incrementally rotates past the seedcollector 114. Alternatively, in various other embodiments, each seedcan be released from respective recessed port 118 by temporarilyterminating the vacuum at each individual recessed port 118 as theindividual recessed port 118 is positioned adjacent the seed collector114. In still other embodiments, each seed can be blown from therespective recessed port 118 by temporarily providing forced air at eachindividual recessed port 118 as the individual recessed port 118 ispositioned adjacent the seed collector 114.

After each seed is removed from the separating wheel 108, the seed isfunneled into one of a plurality of first transfer tubes 126 havingtheir proximal ends connected to openings 128 in a tube shuttle 130. Thetube shuttle 130 is mounted to a carriage 134 that is movably mounted toa linear translation stage 138 that includes an actuator 142controllable by the CCS 700 to bi-directionally move the carriage 134,tube shuttle 130 and proximal ends of the first transfer tubes 126 alongthe translation stage 138. Therefore, as each seed is removed from theseparating wheel 108, the seed is funneled into one of the firsttransfer tubes 126. Then the CCS 700 moves the tube shuttle 130 alongthe translation stage such that a subsequent first transfer tube 126will receive the next seed removed from the separating wheel 108. Thisprocess of removing seeds is repeated until a seed has been depositedinto each of the first transfer tubes 126. As each seed is removed fromthe separating wheel 108 and deposited into a first transfer tube 126,each seed passes through the respective first transfer tuber 126, viagravity, vacuum or forced air, to a queuing stack 150.

As shown in FIG. 3B, the queuing stack 150 includes a plurality of upperchambers 154, e.g., eight upper chambers 154. A distal end of each firsttransfer tube 126 terminates at a corresponding one of the upperchambers 154. Each upper chamber 154 includes an automated upper releasemechanism 156, e.g., a flapper gate, that, under control of the CCS 700,retains the respective seed within the upper chamber 154. Once eachupper chamber has a seed deposited therein, the upper release mechanisms156 are commanded to release the seeds into a plurality, e.g., eight, ofcorresponding lower chambers 158. Similar to the upper chambers 154,each lower chamber 158 includes an automated lower release mechanism160, e.g., a flapper gate, that, under control of the CCS 700, retainsthe respective seed within the lower chamber 154. The lower chambers 158retain the seeds until such time as the CCS 700 commands the lowerrelease mechanisms 160 to release the seeds into a plurality ofcorresponding second transfer tubes 162 having their proximal endsconnected to the lower chambers 158.

As shown in FIG. 3C, a distal end of each second transfer tube 162terminates at a corresponding one a plurality of elevator chambers 164,e.g., eight, of an elevator hopper 166. Thus, as the seeds are releasedfrom the lower chambers 158 into the second transfer tubes 162, eachseed passes through the respective second transfer tube 162, viagravity, vacuum or forced air, into a corresponding one of the elevatorchambers 164. Once a seed is deposed into each elevator chamber 164, thegroup of seeds therein constitutes a set of seeds, as used herein. Theelevator hopper 166 additionally includes a plurality of elevator pistonrods 170. Each elevator piston rod 170 is positioned within, andextendable through, an aperture 174 formed in a funnel-shaped bottom 178of a corresponding elevator chamber 164. In various embodiments, adistal end of each elevator piston rod 170 is formed to have a concaverecess 180 shaped to cradle the seeds received from the lower chambers158 of the queuing stack 150. Additionally, the angled sides of thefunnel-shaped bottom 178 allows for the seeds entering the respectiveelevator chambers 164 to fall onto their sides, i.e., lay flat, and becentered within the recess 180 of the respective elevator piston rods170.

Each elevator piston rod 170 can be extended and retracted through therespective elevator chamber aperture 174 by a corresponding one of aplurality of piston actuators 182, as controlled by the CCS 700. Moreparticularly, prior to the seeds being deposited in the elevatorchambers 164, the CCS 700 commands the piston actuators 182 to retractthe piston rods 170 to a retracted position where the recessed distalends of each piston rod 170 is substantially flush with the bottom ofeach respective elevator chamber 164 within the respective aperture 174,as exemplarily illustrated by the six leftmost piston rods 170 in FIG.3C. Then once the seeds are deposited into the elevator chambers 164,the seeds are retained and cradled within the respective piston rodrecesses 180, until such time as the CCS 700 commands the pistonactuators to extend the piston rods 170 to an extended position, asexemplarily illustrated in phantom by the two rightmost piston rods 170in FIG. 3C. Extending the piston rods 170 raises each respective seedout of the respective elevator chamber 164 to a cued position where theset of seeds are presented for removal, processing and sampling by a RVCbank 204, as describe below.

Referring now to FIGS. 4A and 4B, as described above, the seed transportsubsystem 200 includes a plurality of RVC banks 204 mounted to thetransport carousel 208. For simplicity and clarity, FIG. 4A illustratesa single RVC bank 204 mounted to the transport carousel 208, however, itshould be understood that seed transport subsystem 200 includes aplurality of RVC banks 204 mounted thereto. For example, in variousembodiments, the seed transport subsystem 200 includes four RVC banks204, whereby each RVC bank 204 is mounted to one of four sides of thetransport carousel 208. Each RVC bank includes a plurality of rotaryvacuum cup (RVC) devices 212 controllable by the CCS 700 to remove a setof cued seeds from the elevator piston rods 170 and sequentiallytransport the respective seed set to each of the imaging station 300,the orientation station 400 and the sample and sort station 500.

Each RVC device 212 includes a vacuum cup 216 mounted to a first end ofa rotary shaft 220, and a friction plate 224 mounted to an opposingsecond end of the rotary shaft 220. Each RVC device 212 additionallyincludes a shaft actuator 228 controllable by the CCS 700 tobidirectionally move the shaft 220, vacuum cup 216 and friction plate224 along the longitudinal axis of the rotary shaft 220. That is, eachactuator 228 is controlled by the CCS 700 to raise and lower therespective vacuum cup 216 as needed throughout operation of the seedsorter system 10. Each vacuum cup 216 is communicatively connected to avacuum source (not shown) that is controlled by the CCS 700 toselectively provide a vacuum at a tip 232 of each vacuum cup 216. EachRVC device 212 further includes a biasing device 236, e.g., a spring,configured to apply a constant force on the rotary shaft 220 in the Xdirection. The force applied in the X direction by the biasing devices236 maintains a locking mechanism (not shown) of each respective RVCdevice 212 engaged. Engagement of the locking device prevents angularrotation of the respective rotary shaft 220 and vacuum cup 216 about thelongitudinal axis of the rotary shaft 220 until the locking mechanism isdisengaged, as described below.

In coordination with a set of seeds being loaded into the elevatorchambers 164, the CCS 700 positions an empty RVC bank 204, i.e., an RVCbank 204 without a set of seeds retained by the respective vacuum cups216, above the elevator bank 166. The RVC devices 212 are located andmounted to the transport carousel 208, and the motor of the transportcarousel 208 is controlled, such that when an RVC bank 204 is positionedadjacent the loading station 100, the vacuum cup 216 of each RVC device212 is positioned directly above a corresponding one of the elevatorchambers 164. More particularly, when an RVC bank 204 is positionedadjacent the loading station 100, the vacuum cup 216 of each RVC device212 is positioned directly above the elevator piston rod 170 of thecorresponding elevator chamber 164. Once the elevator chambers 164 areloaded with a set of seeds, and an empty RVC bank 204 is positionedadjacent the loading station 100, the CCS 700 can command the elevatorpiston rods 170 to raise the set of seeds to the cued position. The RVCdevices 212 are further located and mounted to the transport carousel208, such that when the elevator piston rods 170 are in the extendedposition, i.e., the set of seeds are cued, each seed is in light contactwith, or close proximity to, the corresponding vacuum cup 216. A vacuumis then provided to each vacuum cup tip 232. The vacuum cup tips 232 aresized, and fabricated from a suitable material, such that when thevacuum is provided, each respective seed is firmly retained on therespective tip 232. The CCS 700 then retracts the elevator piston rods170 leaving the set of seeds firmly retained on the respective vacuumcup tips 232. The retained set of seeds can then be positioned adjacentthe imaging station 300, via advancement of the transport carousel 204.

Referring now to FIGS. 5A and 5B, in various embodiments, the imagingstation 300 includes at least one imaging device 304 mounted to systemsupport structure such that the one or more imaging devices 304 is/arepositioned under the RVC bank 204 and the respective set of seeds whenthe set of seeds is advanced from the loading station 100. In variousembodiments, the imaging station 300 includes a first imaging device 304positioned and operable to collect image data for a first one-half ofthe seed set, and a second imaging device 304 positioned and operable tocollect image data for a second one-half of the seed set. Moreparticularly, the first imaging device 304 is mounted to the systemsupport structure such that a field of view of the first imaging device304 includes a bottom side of a first half of the seeds positionedadjacent the imaging station 300. And, the second imaging device 304 ismounted to the system support structure such that a field of view of thesecond imaging device 304 includes a bottom side of a second half of theseeds positioned adjacent the imaging station 300.

As used herein, reference to the bottom side of the seeds refers to theside of the seeds that is facing downward with respect to theorientation of each seed as retained by the respective vacuum cup 216.As described above, the shape of the elevator chamber bottoms 178 andthe shape of the recesses 180 at the distal ends of each elevator rod170 are designed such that each seed is preferably retained on thevacuum cup tips 232 by one of the opposing broad sides of eachrespective seed. That is, each seed is preferably held on the respectivevacuum cup 216 by one of the broader sides such that germ of the seed isviewable by the imaging device(s) 304 and the tip of each seed ispointing anywhere within a 360° plane that is substantially orthogonalto the respective vacuum cup 216. The imaging device(s) 304 may be anysuitable imaging device selected in accordance with the imaging goals ofseed sorter system 10. For example, in connection with an analysis forexternal seed coat, the first imaging device 304 may comprise a digitalcamera operable in the visible light range. Alternatively, for internalseed analysis, the first imaging device 304 may comprise a cameraoperable in the near infra-red light range (see, U.S. application patentSer. No. 09/698,214, the disclosure of which is hereby incorporated byreference). Still further, the first imaging device 304 may comprise acamera which implements NMR/MRI imaging techniques (see, U.S.application patent Ser. No. 09/739,871, the disclosure of which ishereby incorporated by reference).

The imaging station 300 further includes a light source 312 mounted tosystem support structure for illuminating the field of view of theimaging device(s) 304. The source 312 can be any type of light sourcesuited for the particular imaging application of the seed sorter system10. For example, the light source 312 can be one or more incandescentlights, fluorescent lights, ultraviolet lights, infrared lights, etc. Invarious embodiments, the light source 312 comprises a bank of lightemitting diodes (LEDs), e.g., 630 nm LEDs. For example, the light source312 comprises a bank of LEDs wherein each seed in the seed set has acorresponding LED as the primary light source illuminating therespective seed. Additionally, in various embodiments, each vacuum cup216 includes a dark colored, e.g., black, background disk 240 (FIG. 4B)that provides a dark background for each seed during imaging andprevents image data interference from system components and structurebeyond the seeds and within the field of view of imaging device(s) 304.

The image data is transmitted to the CCS 700 and stored (at leasttemporarily) in an electronic data storage device of the CCS 700. TheCCS 700 analyzes the data to determine a directional orientation of thetip of each seed. That is, the CCS 700 analyzes the image data todetermine which direction the tip of each individual seed is pointingwithin the 360° plane substantially orthogonal to the respective vacuumcup 216. For example, with reference to FIG. 5B, if a point on the 360°plane that is directly opposite the transfer carousel 208 is consideredthe origin, i.e., 0°, the CCS 700 may analyze the image data todetermine that the tip one of the seeds in the seed set is oriented at90°, while the tip of another of the seeds in the seed set is orientedat 315°, and the tip of yet another seed is oriented at 200°, etc. Oncethe image data of the respective seed set is collected and transmittedto the CCS 700, by the imaging device(s) 304, the transfer carousel 208is advanced to position the seed set adjacent the orientation station400.

Referring now to FIGS. 6A and 6B, in various embodiments, theorientation station 400 includes motor bank 404 that includes aplurality of rotary motors 408, e.g., a number of rotary motors 408equal to the number of RVC 212, independently controlled by the CCS 700.In some embodiments, the rotary motors 408 comprise stepper motors. Eachmotor 408 includes a rotary shaft 412 having a clutch plate 416 mountedto a distal end thereof. The motor bank 404 is mounted to a linearactuator 420, e.g., a pneumatic slide, that is mounted to system supportstructure such that when the RVC bank 204 is positioned adjacent theorientation station 400, each motor 408 is positioned directly above arespective one of the RVC devices 212. More specifically, when the RVCbank 204 is positioned adjacent the orientation station 400, the clutchplate 416 of each motor 408 is positioned directly above, and inalignment with, a respective one of the RVC friction plates 224.

Once the imaged set of seeds is advanced to the orientation station 400,the actuator 420 lowers the motor bank 404 such that the clutch plates416 of each motor 408 engage the corresponding friction plates 224 ofthe respective RVC device 212. Additionally, the actuator 420 is loweredsuch that the clutch plates 416 apply force to each friction plate 224in the Y direction that overcomes the force in the X direction appliedby the RVC biasing devices 236. Accordingly, each RVC friction plate224, rotary shaft 220 and vacuum cup 216 is moved downward, therebydisengaging the RVC locking mechanism and allowing each RVC frictionplate 224, rotary shaft 220 and vacuum cup 216 to rotate. Then, based onthe analyzed image data collected at the imaging station 300, each motor408 is independently controlled by the CCS 700 to rotate the respectivefriction plates 224 and corresponding vacuum cups 216 to independentlyproperly orient each respective seed for sampling at the sample and sortstation 500, as described below. More particularly, based on theanalyzed image data for each independent seed, each motor 408 isindependently controlled to rotate the respective seed such that the tipof the seed is oriented approximately at 0°. More importantly, each seedis independently rotated, if necessary, to position the cap of the seedat approximately 180° such that a sample can be removed from that cap ofeach seed at the sample and sort station 500, as described below.

Once each seed of the set is oriented with the cap of each respectiveseed oriented, or positioned, at approximately 180°, the CCS 700commands the actuator 420 to raise the motor bank 404 to disengage theclutch plates 416 from the friction plates 224. As the motor bank 404 israised and the clutch plates 416 are disengaged from the friction plates224, the biasing devices 236 of each RVC device 212 move each respectiverotary shaft in the X direction thereby engaging each respective lockingdevice. Thus, each rotary cup 216 and corresponding seed held thereon,is maintained in the orientation with the seed cap at approximately the180° position, as illustrated in FIG. 6B. The CCS 700 then advances thetransfer carousel 208 to position the properly oriented seed setadjacent the sample and sort station 500.

Referring now to FIG. 6C, in various embodiments, the orientationstation 400 further includes a seed purge hopper 424 for receiving theset of seeds held by the respective RVC bank 204. The seed purge hopper424 is mounted to system support structure such that a trough 428 of theseed purge hopper 424 is positioned under the vacuum cups 216 of therespective RVC bank 204 for receiving seeds discharged from therespective vacuum cups 216. More specifically, the seed purge hopper 424can be utilized to offload all the seeds held by each RVC bank 204 ofthe seed transport subsystem 200. To offload all the seeds, each RVCbank 204 is sequentially advanced to the orientation station 400 atwhich time the vacuum source being supplied to each respective vacuumcup 216 is terminated. When the supplied vacuum is terminated, the seedsare released from the vacuum cups 216 and fall into the seed purgehopper trough 428 where they can be collected and returned to the bulkseed hopper 104 at a later time. Thus, if operation of the seed sortingsystem 10 needs to be terminated, all the seeds held by the RVC banks204 can be purged and collected.

In various embodiments, the seed sorting system 10 includes an emergencystop button 22, shown in FIG. 1, for stopping and shutting down the seedsorting system 10. For example, in the case of an emergency, theemergency stop button 22 can be depressed and the all operation of theseed sorting system 10 would cease. Also, in various embodiments, theseed sorting system 10 includes a system pause button 26, shown in FIG.1, for temporarily pausing operation of the seed sorting system 10. Forexample, if a jam occurred in one of the first transfer tubes 126 of theseed loading station 100 such that one or more RVC banks did not ‘pickup’ a full set of seeds, the system pause button could be depressed topause operation of the seed sorting system 10. In the paused state, thevacuum source can remain actuated such that all seeds are retained bythe respective RVC vacuum cups 216 until such time as the seeds arepurged into the seed purge hopper 424 or operation is reinitiated andthe seeds are sampled, as described below.

Referring now to FIG. 7, in various embodiments, the seed sample andsort station 500 includes a seed sampling subsystem 510 and a seed andsample sorting subsystem 570. The seed sampling subsystem 510 iscontrollable by the CCS 700 to extract a sample from each seed in therespective seed set positioned adjacent the seed sample and sort station500. The seed and sample sorting subsystem 570 is additionallycontrollable by the CCS 700 to sort the sampled seeds to the seed trays18 and sort the corresponding seed sample to the sample trays 14 whiletracking and mapping the locations of the corresponding sampled seedsand seed samples in the respective seed and sample trays 18 and 14. Thelocations of the seed samples and the locations of the correspondingsampled seeds in the trays 14 and 18 are matched so that the sampledseed may later be correlated to the sample taken therefrom (e.g., afteranalysis of the sample, etc.).

Referring now to FIGS. 8A, 8B, 8C and 8D, in various embodiments, theseed sampling subsystem 510 includes a plurality of seed grip and chipassemblies 512, e.g., a number of seed grip and chip assemblies 512equal to the number of RVC devices 212 included in each RVC bank 204.The seed sampling subsystem 510 additionally includes a press plate bank514 mounted to a linear actuator 516, e.g., a pneumatic slide. The pressplate bank 514 includes a plurality of press plates 518 fixedly mountedto a press plate bank header 520 that is coupled to the linear actuator516. The actuator 516 is mounted to system support structure such thatwhen the RVC bank 204 is positioned adjacent the sample and sort station500, each press plate 518 is positioned directly above a respective oneof the RVC devices 212. More specifically, when the RVC bank 204 ispositioned adjacent the orientation station 400, each press plate 518 ispositioned directly above, and in alignment with, a respective one ofthe RVC friction plates 224.

Once the oriented set of seeds is advanced to the sample and sortstation 500, the actuator 516 lowers the push plate bank 514 such thatthe push plates 518 engage the corresponding friction plates 224 of therespective RVC device 212. As the actuator 420 is lowered, the pushplates 518 apply force to each friction plate 224 in the Y directionthat overcomes the force in the X direction applied by the RVC biasingdevices 236. Accordingly, each RVC friction plate 224, rotary shaft 220and vacuum cup 216 is moved downward, thereby disengaging the RVClocking mechanism. However, since the press plates 518 are fixedlymounted to the header 520, each RVC friction plate 224, rotary shaft 220and vacuum cup 216 can not rotate and each seed remains properlyoriented as it is moved downward in the Y direction.

With particular reference to FIGS. 8B and 8D, in accordance with variousembodiments, each grip and chip assembly 512 includes a seed grippingmechanism 522 and a sample extraction mechanism 524. Although, each gripand chip assembly 512 is independently controlled by the CCS 700, thestructure and function for each grip and chip assembly 512 issubstantially identical. Therefore, the structure and function of theplurality of grip and chip assemblies will be described herein withreference to a single grip and chip assembly 512. The seed grippingmechanism 522 is operable, as controlled by the CCS 700, to firmly holdeach respective seed as the sample extraction mechanism 524 removes aportion, i.e., a sample, of the seed coat and inner seed material fromthe crown of the respective seed. The extracted sample can then beutilized to test and analyze the various traits of the respective seed.Importantly, the sample is extracted from the crown in a non-destructivemanner such that germination viability of the seeds can be preserved.

In various embodiments, the seed gripping mechanism 522 includes anactuator 526, e.g., a pneumatic clamp, that is controllable by the CCS660 to bidirectionally move a pair of opposing actuator arms 528 towardand away from each other, i.e., open and close the actuator arms 528.For example, in various embodiments, the actuator 526 is operable tomove the opposing actuator arms 528 toward and away from each otheralong the line M (FIG. 8B). The actuator arms 528 are structured toremovably retain a pair of opposing seed gripping fingers 530 structuredto firmly hold the respective seed as the sample is extracted by thesample extraction mechanism 524, as described below. An exemplarygripping finger 530 is illustrated in FIG. 8C. Each gripping finger 530includes a head 532 having a contoured face 534. The face 534 can beshaped or formed to have any conformation suitable for firmly andsteadily holding the respective seed as the sample is extracted. Invarious embodiments, the face 534 is particularly designed to have awedge-like conformation such that as the actuator 526 closes grippingfingers 530 around the seed, the seed is forced toward a cutting wheel540 of the sample extraction mechanism 524 and into abutment with ajustification block 562 of the sample extraction mechanism 524. Thejustification block 562 includes a cutting wheel guide slot 533 thatallows the cutting wheel 540 access to seed. Thus, the gripping fingers530 firmly and justification block 562 hold the seed on three sides andprevent the respective seed from moving in a direction away from thesample extraction mechanism 524 as the sample is being extracted. Invarious embodiments, the gripping finger head 532 is connected to, orintegrally formed with, a mounting post 536 structured to fit within, ormate with, a mounting hole 538 in each actuator arm 528.

When the RVC bank 204 and properly oriented seed set are advanced to theseed sample and sort station 500, the CCS 700 commands the seed grippingactuator 526 to open the actuator arms 528 such that the gripping fingerfaces 534 have a space between them large enough to allow a seed to beeasily positioned therebetween. The CCS 700 then commands the pressplate bank actuator 516 to lower the press plate bank 514 to engage thepress plates 518 with the friction plates 224. More particularly, theforce on the friction plates 224 moves the vacuum cups 216 andrespective seed downward toward a sampling position, i.e., the gapbetween gripping fingers 530. Each grip and chip system 512 is mountedto system support structure such that each sampling position, or gap,between the gripping finger faces 534 is precisely aligned below therespective vacuum cup tip 232. Thus, when the press plate bank actuator516 pushes the friction plates 224, vacuum cups 216 and oriented seedsdownward, the oriented seeds are moved to the sampling positions betweenthe gripping fingers 530 of the respective seed gripping mechanism 522.

The CCS 700 then commands the seed gripping actuator 526 to close theactuator arms 528 such that the gripping finger faces 534 engage therespective seed to firmly retain the seed without damaging the seed.Once the seed is firmly retained between the gripping fingers 530, thepush plate bank actuator can be commanded to raise, and the vacuumprovided at the vacuum cup tip 232 terminated, to thereby release therespective seed. Or, alternatively, the CCS 700 can maintain the vacuumcup 216 in contact with the seed to provide additionally support for theseed as the sample is being extracted.

The sample extraction mechanism 524 includes cutting wheel 540rotationally mounted within a cutting wheel fixture 542 and rotationallydriven by a cutting wheel motor 544. Although the cutting wheel 540 isshown in FIG. 8B to be belt driven by the cutting wheel motor 544,alternatively the cutting wheel 540 can be shaft driven, chain driven,direct gear driven, etc., by the cutting wheel motor 544 and remainwithin the scope of the present disclosure. The cutting wheel 540 ismounted on a shaft 546 that is rotationally mounted within the cuttingwheel fixture 542. Additionally, a drive wheel 548 is mounted to, orformed with, the shaft and operatively coupled to the cutting wheelmotor 544, for example, by a drive belt 550, such that actuation of themotor 544 will rotate the drive wheel 548, shaft 546 and cutting wheel540 within the cutting wheel fixture 542. More specifically, the cuttingwheel 540 is mounted to the shaft 546 in a cam fashion, e.g., the shaft546 can be an offset shaft, such that as the drive wheel 548 and shaft546 are rotated by the motor 544, a peripheral cutting edge 552 of thecutting wheel 540 rotates and progressively moves toward the seedgripping mechanism 552, and specifically toward the seed retainedbetween the gripping fingers 530. The cutting edge 552 comprises anabrasive or sharp-edged surface, e.g., a saw-toothed surface, that willremove the seed coat and inner seed material from the crown of therespective seed. Thus, as the cutting wheel 540 is rotated, the cuttingedge 552 will contact and begin to cut or abrade the seed crown. As thecutting wheel 540 continues to rotate, the cutting edge 552 willpenetrate a desired depth or distance into the crown, depending on theamount of angular rotation of the cutting wheel 540, as controlled bythe CCS 700. That is, the greater the amount of angular rotation of thecutting the wheel 540, the deeper the cutting edge 552 will penetrateinto the seed crown and the more sample that will be extracted.

The seed gripping mechanism 522 additionally includes a seed dump bowl554 (FIG. 8D) having a drain port 556 formed in the bottom of the dumpbowl 554. The dump bowl 554 is shaped to funnel a sampled seed into thedrain port 556 after a sample has been removed from the respective seedand the seed is released from being held between the gripper fingers530. A drain tube 558 (shown in FIGS. 9A and 9B) is connected to thedrain port 556 to transfer the released seed to one of the seed trays 18positioned below the grip and chip assembly 512, as describe below. Theseed gripping mechanism 522 further includes a sample extraction orifice560 located in the justification block 562, below the cutting wheelguide 533, such that cutting edge 552 and periphery portion of thecutting wheel 540 extends over the sample extraction orifice 560. Asample extraction tube 564 (FIGS. 9A and 9B) is connected to the sampleextraction orifice 560 and a vacuum is controllably provided to thesample extraction tube 564 and thus, at the sample extraction orifice560. As the cutting wheel 540 removes the sample from the respectiveseed, the vacuum provided at the sample extraction orifice 560, via thesample extraction tube 564, draws the sample into the sample extractionorifice 560. The sample is then passed through the sample extractiontube 564 and deposited into one of the sample trays 14.

Therefore, once the seed is retained between the gripping fingers 530,the CCS 700 commands the cutting wheel motor 544 to angularly rotate thedrive wheel 548 through a predetermined angle and at a predeterminedrate of rotation. For example, the CCS 700 can command the cutting wheelmotor 554 to rotate the drive wheel 548 ninety-five degrees at thirtyrevolutions-per-minute (RPMs). Accordingly, the cutting wheel 540 isrotated through the predetermined angle, at the predetermined RPMs. Asthe cutting wheel 540 rotates, the cam action of cutting wheel 540mounting rotates and advances the cutting edge 552 toward and into theseed, thereby removing a sample from the respective seed. As the sampleis removed, the vacuum at the sample extraction orifice 560 draws thesample into the sample extraction tube 564 where the sample istransferred to one of the sample trays 14. The CCS 700 then commands thecutting wheel motor 544 to reverse the direction of rotation to withdrawthe cutting wheel 540 from the seed and return the cutting wheel to ahome position, ready to remove a sample from a subsequent seed.Subsequent to, or substantially simultaneously with the withdrawal ofthe cutting wheel 540, the CCS 700 commands the seed gripping mechanism522 to release the sampled seed, allowing the seed to fall, via gravity,vacuum or forced air, into the drain tube 556 to transfer the sampledseed to one of seed trays 18.

Referring now to FIG. 8E, in various embodiments, the cutting wheel 540is structured to have a saw-toothed cutting edge 552 that includes aplurality of teeth 566. Moreover, each tooth 566 includes a lateralcutting tip 568 that is formed to avoid movement, e.g., ‘chattering’, ofthe seed being sampled and allow the seed to remain stationary withinthe gripping fingers 530. For example, the lateral cutting tip 568 ofeach tooth can have a specific angle α, e.g, a 60° angle, such that asthe cutting wheel 540 cuts through the respective seed, a leading end ofthe cutting tip 568 of each subsequent tooth 566 engages the seed beforea trailing end of the cutting tip 568 of each preceding tooth 566disengages the seed.

In various embodiments, each cutting wheel 540, i.e., each cutting wheelmotor 544, is independently controlled by the CCS 700, but each cuttingwheel 540 is commanded to have approximately the same rotational speedand/or angular rotation. Therefore, when a set of seeds is held withinthe seed gripping mechanisms 522, the respective cutting wheels 540 areeach commanded to rotate through approximately the same angle ofrotation and at the same speed. Accordingly, the cam rotation of thecutting wheels 540, as described above, will advance the respectivecutting wheels 540 approximately the same distance toward each of theseed. Thus, smaller seeds may not have the same amount of sampleextracted as larger seeds. In such embodiments, the rotational speedand/or amount of angular rotation for each cutting wheel 540 isdetermined by empirical data and programmed into the CCS 700.

In various other embodiments, each cutting wheel 540, i.e., each cuttingwheel motor 544, is independently controlled by the CCS 700. Therefore,the rotational speed and/or amount of angular rotation for eachindependent cutting wheel 540 can be controlled and adjusted for eachseed positioned and held by the seed gripping mechanism 522 of eachrespective grip and chip assembly 512. For example, the seed held withina seed gripping mechanism 522 of a first grip and chip assembly 512 maybe smaller in size than a seed held within a seed gripping mechanism 522of an adjacent second grip and chip assembly 512. In such a case, thecutting wheel 540 of first grip and chip assembly 512 can be commandedto have a greater angular rotation than the cutting wheel 540 of secondgrip and chip assembly 512. Therefore, the cam rotation of the cuttingwheels 540, as described above, will advance the cutting wheel 540 ofthe first chip and grip assembly 512 further toward the smaller seedsuch that approximately equal amounts of sample will be extracted fromthe smaller seed as from the larger seed. Furthermore, in suchembodiments, the rotational speed and/or amount of angular rotation foreach independent cutting wheel is based on the imaging data collectedfor each respective seed at the imaging station 300.

Referring now to FIGS. 9A, 9B, 9C, 9D and 9E, as described above, as thesample is extracted from the respective seed, the sample is drawn intothe sample extraction tube 564. More specifically, the sample extractionorifice 560 of each grip and chip assembly 512 has a first end of arespective sample extraction tube 564 connected thereto, and a secondend of each respective sample extraction tube 564 is connected to asample extraction nozzle (SEN) manifold 572. The SEN manifold 572includes a manifold block 574 to which the sample extraction tubes 564are connected, and from which a plurality of exhaust tubes 578 extend,e.g., a number of exhaust tubes 578 equal to the number of sampleextraction tubes 564 can extend from the manifold block 574. The SENmanifold 572 additionally includes a plurality of discharge nozzles 580that are in fluid communication with the extraction tubes 564. Morespecifically, the manifold block 574 includes a number of bores orpassages (not shown), equal to the number of sample extraction tubes564, which extend through the manifold block 574. Each extraction tube564 is connected to a first end of a corresponding manifold block boreand a corresponding one of the discharge nozzles 580 extends from anopposing second end of each manifold block bore.

As most clearly illustrated in FIG. 9D, the seed and sample sortingsubsystem 570 further includes a sample tray platform 582 adapted tosecurely retain a plurality of the sample trays 14 in fixed positionsand orientations. Each sample tray 14 includes a plurality of samplewells 30, each of which are adapted for receiving a sample extracted byone of the grip and chip assemblies 512. For example, in variousembodiments, each sample tray 14 can be a ninety-six well tray.Moreover, the discharge nozzles 580 extending from the SEN manifold 572are spaced apart and arranged to be congruent with the spacing andarrangement of the wells 30 within the sample trays 14. The sample trayplatform 582 is mounted to an X-Y stage 584 that is a two-dimensionaltranslation mechanism, including a X axis translating track 586 and a Yaxis translating track 588. The X-Y stage 584 additionally includes afirst linear actuator 590 operable to bidirectionally move a firstcarriage (not shown) along the length of the X axis translating track586. The X-Y stage 584 further includes a second linear actuator 592operable to bidirectionally move a second carriage (not shown) along thelength of the Y axis translating track 588. The Y axis translating track588 is mounted to the first carriage and the sample tray platform 582 ismounted to the second carriage.

The SEN manifold 572 is connected to system support structure toposition the SEN manifold 572 above the X-Y stage 584 and the sampleplatform 582 holding the plurality of sample trays 14. Moreparticularly, the SEN manifold 572 is mounted to a linear actuator 594,e.g., a pneumatic slide, controllable by the CCS 700 to bidirectionallymove the SEN manifold 572 along the Z axis, e.g., up and down. The firstand second linear actuators 590 and 592 are controlled by the CCS 700 toprecisely move the sample tray platform 582 in two dimensions. Moreparticularly, the first and second actuators 590 and 592 move the sampletray platform 582 within an X-Y coordinate system to precisely positionany selected group of adjacent wells 30 of any one or more selectedsample trays 14 at a target location directly beneath the SEN manifold572.

In operation, prior to the grip and chip assemblies 512 extractingsamples from the respective seeds advanced to the seed sample and sortstation 500, the CCS 700 controls the X-Y stage 584 to position aselected group of wells 30 at the target location. The CCS 700 thencommands the SEN manifold actuator 594 to lower the SEN manifold 572 toposition each of the discharge nozzles 580 in alignment with and inclose proximity to, or in contact with, a corresponding one of the wells30. Once the selected group of wells 30 is positioned at the targetlocation and the SEN manifold 572 is lowered, the CCS 700 commands thegrip and chip assemblies 512 to extract the samples from the respectiveseeds. Each sample is drawn into a corresponding sample extraction tube564, as described above, and the vacuum provided to each sampleextraction tube 564 transfers each sample through the respective sampleextraction tube 564 to the corresponding discharge nozzle 580. Each seedis then discharged into the corresponding sample tray wells 30. The SENmanifold actuator 594 then raises the SEN manifold 572, a subsequentgroup of wells 30 is positioned at the target position, and the SENmanifold 572 is lowered in preparation for a subsequent set of samplesto be extracted and deposited into the wells 30.

In various embodiments, each discharge nozzle 580 includes a seal 596that contacts the sample tray(s) 14 and creates a seal between eachdischarge nozzle 580 and the corresponding well 30 when the SEN manifold572 is lowered. Thus, the seals 596 ensure that substantially all thesample being discharged from each discharge nozzle 580 is deposited intothe corresponding well 30 without cross-contamination by adjacentsamples escaping around the discharge nozzles 580. The seals 596 can beany seal suitable for creating a seal between each discharge nozzle 580and the corresponding well 30, e.g., an O-ring, gasket or bushing.

As the sample trays 14 are placed on the sample tray platform 582, atray identification number, e.g., a bar code, for each sample tray 14and the location of each sample tray 14 on the platform 582 is recorded.Additionally, as each extracted sample is deposited into a well 30, anX-Y location of the well 30 on the sample tray platform 582 is recorded.The recorded tray and well positions on the sample tray platform 582 canthen be compared to the X-Y locations of each deposited extractedsample, to map the specific extracted sample in each well 30 of eachsample tray 14. In various embodiments, the sample tray platform 582 isremovably coupled to the X-Y stage 584 such that one or more sample trayplatforms 582 can be loaded with the sample trays 14 offline, i.e., awayfrom the seed sorter system 10, and conveniently coupled to anddecoupled from the X-Y stage 584.

Additionally, in various embodiments, the extraction tubes 564 arefabricated from static dissipative tubing so that a portion of theextracted samples do not stick to the inside walls of the extractiontubes 564 and cause cross-contamination of the samples. Furthermore, invarious embodiments, the seed and sample sorting subsystem is structuredand operable to ‘blow out’ the extraction tubes 564 and dischargenozzles 580 between cycles. Therefore, any sample residue accumulated inthe extraction tubes 564 and discharge nozzles 580 is cleaned outbetween cycles. For example, air pressure can be drawn or forced throughthe extraction tubes 564 and discharge nozzles 580 and exhausted intothe exhaust tubes 578. The exhaust tubes 578 can be coupled to anexhaust manifold 598 that carries any residual sample particles tocollection chambers 600, where the particles are filtered out of theexhausted air, i.e., separated from the exhausted air.

As most clearly illustrated in FIG. 9E, the seed and sample sortingsubsystem 570 still further includes a seed tray platform 602 adapted tosecurely retain a plurality of the seed trays 18 in fixed positions andorientations. Each seed tray 18 includes a plurality of seed wells 34,each of which are adapted for receiving a seed after the respective seedhas been sampled by one of the grip and chip assemblies 512. Forexample, in various embodiments, each seed tray 18 can be a twenty-fourwell tray. The bank of grip and chip assemblies 512 is mounted to systemsupport structure above the seed tray platform 602 such that seeds canbe dispensed through the drain tubes 558 into selected seed wells 30 ofselected seed trays 18.

The seed tray platform 602 is mounted to an X-Y stage 604. The X-Y stage604 is a two-dimensional translation mechanism, including an X axistranslating track 606 and a Y axis translating track 608. The X-Y stage604 additionally includes a first linear actuator 610 operable tobidirectionally move a first carriage (not shown) along the length ofthe X axis translating track 606. The X-Y stage 604 further includes asecond linear actuator 612 operable to bidirectionally move a secondcarriage (not shown) along the length of the Y axis translating track608. The Y axis translating track 608 is mounted to the first carriageand the seed tray platform 602 is mounted to the second carriage.

The first and second linear actuators 610 and 612 are controlled by theCCS 700 to precisely move the seed tray platform 602 in two dimensions.More particularly, the first and second actuators 610 and 612 move theseed tray platform 602 within an X-Y coordinate system to preciselyposition any selected well 34 of any selected seed tray 18 at a targetlocation beneath a selected one or more of the drain tubes 558. Invarious embodiments, the drain tubes 558 are held in linear alignment bysystem support structure and the CCS 700 controls the first and secondactuators 610 and 612 to position a selected group of linearly adjacentwells 34 at a target location beneath the linearly aligned drain tubes558. More specifically, the CCS 700 moves the seed tray platform 602within the X-Y coordinate system to position each of a plurality oflinearly adjacent wells 34 beneath a corresponding one of the linearlyaligned drain tubes 558. Therefore, when each of the seed grippingmechanisms 522 releases the respective sampled seeds, each sampled seedwill fall, via gravity, vacuum or forced air, through the respectivedrain tube 558 into the corresponding well 34 located beneath therespective drain tube 558.

In operation, just prior to, simultaneously with, or just after the setof seeds is retained by the seed gripping mechanisms 522, as describedabove, the CCS 700 positions the selected well 34, or selected group ofwells 34, at the target location. Each seed is then sampled and thesamples are deposited in the sample tray wells 30, as described above.Each seed gripping mechanism 522 is commanded to release the respectiveseeds allowing the seeds to fall into the respective seed dump bowls554. Each seed dump bowl 554 funnels the respective seeds through therespective drain port 556 and into the respective drain tubes 558. Thedrain tubes 558 then direct the respective seeds into the selected wells34 positioned below the drain tubes 558. In various embodiments, the oneor more of the seeds can be sequentially released and the seed trayplatform 602 sequentially repositioned to deposit the one or more seedsinto selected wells 34. In other embodiments, the seed tray platform ismanipulated to position a group of linearly adjacent wells beneath thelinearly aligned drain tubes, all the seeds are then substantiallysimultaneously released and deposited into the respective group oflinearly adjacent wells.

As the seed trays 18 are placed on the seed tray platform 602, a trayidentification number, e.g., a bar code, for each seed tray 18 and thelocation of each seed tray 18 on the seed tray platform 602 is recorded.Additionally, as each seed is deposited in a well 34, an X-Y location ofthe well on the seed tray platform 602 can be recorded. The recordedtray and well positions on the seed tray platform 602 can then becompared to the X-Y locations of each deposited seed, to map thespecific seed in each well 34 of each seed tray 18. In variousembodiments, the seed tray platform 602 is removably coupled to the X-Ystage 604 such that one or more seed tray platforms 602 can be loadedwith the seed trays 18 offline, i.e., away from the seed sorter system10, and conveniently coupled to and decoupled from the X-Y stage 604.

As described above, each of the seed trays 18 and the sample trays 14include a plurality of wells 34 and 30, respectively. In variousembodiments, the number and arrangement of the wells 34 in the seedtrays 18 corresponds to the number and arrangement of the wells 30 inthe sample trays 14. This facilitates a one-to-one correspondencebetween a seed and its extracted sample.

As described above, the sampling systems and methods of this disclosureprotect germination viability of the seeds so as to be non-destructive.Germination viability means that a predominant number of sampled seeds(i.e., greater than 50% of all sampled seeds) remain viable aftersampling. In some particular embodiments, at least about 75% of sampledseeds, and in some embodiments at least about 85% of sampled seedsremain viable. It should be noted that lower rates of germinationviability may be tolerable under certain circumstances or for certainapplications, for example, as genotyping costs decrease with timebecause a greater number of seeds could be sampled for the same genotypecost.

In yet other embodiments, germination viability is maintained for atleast about six months after sampling to ensure that the sampled seedwill be viable until it reaches the field for planting. In someparticular embodiments, the methods of the present disclosure furthercomprise treating the sampled seeds to maintain germination viability.Such treatment may generally include any means known in the art forprotecting a seed from environmental conditions while in storage ortransport. For example, in some embodiments, the sampled seeds may betreated with a polymer and/or a fungicide to protect the sampled seedwhile in storage or in transport to the field before planting.

In various embodiments, the samples of the present disclosure are usedin a high-throughput, non-destructive method for analyzing individualseeds in a population of seeds. The method comprises removing a samplefrom the seed while preserving the germination viability of the seed;and screening the sample for the presence or absence of one or morecharacteristics indicative of a genetic or chemical trait. The methodmay further comprise selecting seeds from the population based on theresults of the screening; and cultivating plants from the selected seed.

Although the present disclosure exemplarily describes thehigh-throughput sampling of maize seeds, one skilled in the art wouldrecognize that any seed can generally be utilized in a method or deviceof the present invention. For example, in various embodiments, the seedcan be selected from the group consisting of alfalfa seed, apple seed,banana seed, barley seed, broccoli seed, cabbage seed, canola seed,carrot seed, castorbean seed, cauliflower seed, Chinese cabbage seed,citrus seed, clover seed, coconut seed, coffee seed, maize seed, cottonseed, cucumber seed, Douglas fir seed, dry bean seed, eggplant seed,Eucalyptus seed, fennel seed, garden bean seed, gourd seed, leek seed,lettuce seed, Loblolly pine seed, linseed seed, melon seed, oat seed,okra seed, olive seed, onion seed, palm seed, pea seed, peanut seed,pepper seed, poplar seed, pumpkin seed, Radiata pine seed, radish seed,rapeseed seed, rice seed, rye seed, spinach seed, sorghum seed, squashseed, Southern pine seed, soybean seed, strawberry seed, sugarbeet seed,sugarcane seed, sunflower seed, sweet corn seed, sweetgum seed, teaseed, tobacco seed, tomato seed, turf seed, watermelon seed, wheat seed,and Arabidopsis thaliana seed. In a more particular embodiment, the seedis selected from the group consisting of cotton seed, cucumber seed,maize seed, soybean seed, rapeseed seed, rice seed, okra seed,watermelon seed and wheat seed. In an even more particular embodiment,the seed is a maize seed, a cotton seed, a cucumber seed or a watermelonseed.

DNA may be extracted from the sample using any DNA extraction methodsknown to those of skill in the art which will provide sufficient DNAyield, DNA quality, and PCR response. A non-limiting example of suitableDNA-extraction methods is SDS-based extraction with centrifugation. Inaddition, the extracted DNA may be amplified after extraction using anyamplification method known to those skilled in the art. For example, onesuitable amplification method is the GenomiPhi® DNA amplification prepfrom Amersham Biosciences.

The extracted DNA is screened for the presence or absence of a suitablegenetic marker. A wide variety of genetic markers are available andknown to those of skill in the art. The DNA screening for the presenceor absence of the genetic marker can be used for the selection of seedsin a breeding population. The screening may be used to select for QTL,alleles, or genomic regions (haplotypes). The alleles, QTL, orhaplotypes to be selected for can be identified using newer techniquesof molecular biology with modifications of classical breedingstrategies.

In other various embodiments, the seed is selected based on the presenceor absence of a genetic marker that is genetically linked with a QTL.Examples of QTLs which are often of interest include but are not limitedto yield, lodging resistance, height, maturity, disease resistance, pestresistance, resistance to nutrient deficiency, grain composition,herbicide tolerance, fatty acid content, protein or carbohydratemetabolism, increased oil content, increased nutritional content, stresstolerance, organoleptic properties, morphological characteristics, otheragronomic traits, traits for industrial uses, traits for improvedconsumer appeal, and a combination of traits as a multiple trait index.Alternatively, the seed can be selected based on the presence or absenceof a marker that is genetically linked with a haplotype associated witha QTL. Examples of such QTL may again include, without limitation,yield, lodging resistance, height, maturity, disease resistance, pestresistance, resistance to nutrient deficiency, grain composition,herbicide tolerance, fatty acid content, protein or carbohydratemetabolism, increased oil content, increased nutritional content, stresstolerance, organoleptic properties, morphological characteristics, otheragronomic traits, traits for industrial uses, traits for improvedconsumer appeal, and a combination of traits as a multiple trait index.

Selection of a breeding population could be initiated as early as the F₂breeding level, if homozygous inbred parents are used in the initialbreeding cross. An F₁ generation could also be sampled and advanced ifone or more of the parents of the cross are heterozygous for the allelesor markers of interest. The breeder may screen an F₂ population toretrieve the marker genotype of every individual in the population.Initial population sizes, limited only by the number of available seedsfor screening, can be adjusted to meet the desired probability ofsuccessfully identifying the desired number of individuals. See Sedcole,J. R. “Number of plants necessary to recover a trait.” Crop Sci.17:667-68 (1977). Accordingly, the probability of finding the desiredgenotype, the initial population size, and the targeted resultingpopulation size can be modified for various breeding methodologies andinbreeding level of the sampled population.

The selected seeds may be bulked or kept separate depending on thebreeding methodology and target. For example, when a breeder isscreening an F₂ population for disease resistance, all individuals withthe desired genotype may be bulked and planted in the breeding nursery.Conversely, if multiple QTL with varying effects for a trait such asgrain yield are being selected from a given population, the breeder maykeep individual identity preserved, going to the field to differentiateindividuals with various combinations of the target QTL.

Several methods of preserving single seed identity can be used whiletransferring seed from a sampling facility to the field. Methodsinclude, but are not limited to, transferring selected individuals toseed tape, a cassette tray, or indexing tray, transplanting with peatpots, and hand-planting from individual seed packets. Multiple cycles ofselection can be utilized depending on breeding targets and geneticcomplexity.

The screening methods of the disclosure may further be used in abreeding program for introgressing a trait into a plant. Such methodscomprise removing a sample comprising cells with DNA from seeds in apopulation, screening the DNA extracted from each seed for the presenceor absence of at least one genetic marker, selecting seeds from thepopulation based upon the results of the DNA screening; cultivating afertile plant from the seed; and utilizing the fertile plant as either afemale parent or male parent in a cross with another plant.

Examples of genetic screening to select seeds for trait integrationinclude, without limitation, identification of high recurrent parentallele frequencies, tracking of transgenes of interest or screening forthe absence of unwanted transgenes, selection of hybrid testing seed,and zygosity testing.

The identification of high recurrent pair allele frequencies via thescreening methods of the present disclosure again allows for a reducednumber of rows per population and an increased number of populations, orinbred lines, to be planted in a given field unit. Thus, the screeningmethods of the present disclosure may also effectively reduce theresources required to complete the conversion of inbred lines.

The methods of the present disclosure further provide quality assurance(QA) and quality control by assuring that regulated or unwantedtransgenes are identified and discarded prior to planting.

The methods of the present disclosure may be further applied to identifyhybrid seed for transgene testing. For example, in a conversion of aninbred line at the BCnF₁ stage, a breeder could effectively create ahybrid seed lot (barring gamete selection) that was 50% hemizygous forthe trait of interest and 50% homozygous for the lack of the trait inorder to generate hybrid seed for testing. The breeder could then screenall F₁ seeds produced in the test cross and identify and select thoseseeds that were hemizygous. Such method is advantageous in thatinferences from the hybrid trials would represent commercial hybridgenetics with regard to trait zygosity.

Other applications of the screening methods of this disclosure foridentifying and tracking traits of interest carry the same advantagesidentified above with respect to required field and labor resources.Generally, transgenic conversion programs are executed in multi-seasonlocations which carry a much higher land and management cost structure.As such, the impact of either reducing the row needs per population orincreasing the number of populations within a given field unit aresignificantly more dramatic on a cost basis versus temperateapplications.

Still further, the screening methods of this disclosure may be used toimprove the efficiency of the doubled haploid program through selectionof desired genotypes at the haploid stage and identification of ploidylevel to eliminate non-haploid seeds from being processed and advancingto the field. Both applications again result in the reduction of fieldresources per population and the capability to evaluate a larger numberof populations within a given field unit.

In various embodiments, the disclosure further provides an assay forpredicting embryo zygosity for a particular gene of interest (GOI). Theassay predicts embryo zygosity based on the ratio of the relative copynumbers of a GOI and of an internal control (IC) gene per cell or pergenome. Generally, this assay uses an IC gene that is of known zygosity,e.g., homozygous at the locus (two IC copies per diploid cell), fornormalizing measurement of the GOI. The ratio of the relative copynumbers of the IC to the GOI predicts the GOI copy number in the cell.In a homozygous cell, for any given gene (or unique genetic sequence),the gene copy number is equal to the cell's ploidy level since thesequence is present at the same locus in all homologous chromosomes.When a cell is heterozygous for a particular gene, the gene copy numberwill be lower than the cell's ploidy level. The zygosity of a cell atany locus can thus be determined by the gene copy number in the cell.

In some particular embodiments, the disclosure provides an assay forpredicting corn embryo zygosity. In corn seed, the endosperm tissue istriploid, whereas the embryo tissue is diploid. Endosperm that ishomozygous for the IC will contain three IC copies. Endosperm GOI copynumber can range from 0 (homozygous negative) to 3 (homozygouspositive); and endosperm GOI copy number of 1 or 2 is found in seedheterozygous for the GOI (or hemizygous for the GOI if the GOI is atransgene). Endosperm copy number is reflective of the zygosity of theembryo: a homozygous (positive or negative) endosperm accompanies ahomozygous embryo, heterozygous endosperm (whether a GOI copy number of1 or 2) reflects a heterozygous (GOI copy number of 1) embryo. Theendosperm GOI copy number (which can range from 0 to 3 copies) can bedetermined from the ratio of endosperm IC copy number to endosperm GOIcopy number (which can range from 0/3 to 3/3, that is, from 0 to 1),which can then be used to predict zygosity of the embryo.

Copy numbers of the GOI or of the IC can be determined by any convenientassay technique for quantification of copy numbers, as is known in theart. Examples of suitable assays include, but are not limited to, RealTime (TaqMan®) PCR (Applied Biosystems, Foster City, Calif.) andInvader® (Third Wave Technologies, Madison, Wis.) assays. Preferably,such assays are developed in such a way that the amplificationefficiency of both the IC and GOI sequences are equal or very similar.For example, in a Real Time TaqMan® PCR assay, the signal from asingle-copy GOI (the source cell is determined to be heterozygous forthe GOI) will be detected one amplification cycle later than the signalfrom a two-copy IC, because the amount of the GOI is half that of theIC. For the same heterozygous sample, an Invader® assay would measure aGOI/IC ratio of about 1:2 or 0.5. For a sample that is homozygous forboth the GOI and the IC, the GOI signal would be detected at the sametime as the IC signal (TaqMan®), and the Invader assay would measure aGOI/IC ratio of about 2:2 or 1.

These guidelines apply to any polyploid cell, or to haploid cells (suchas pollen cells), since the copy number of the GOI or of the IC remainproportional to the genome copy number (or ploidy level) of the cell.Thus, these zygosity assays can be performed on triploid tissues such ascorn endosperm.

The description herein is merely exemplary in nature and, thus,variations that do not depart from the gist of that which is describedare intended to be within the scope of the teachings. Such variationsare not to be regarded as a departure from the spirit and scope of theteachings.

What is claimed is:
 1. An automated method for operating an automatedseed sampling system having a seed loading station, a seed transportsubsystem, and a seed sampling station, the method comprising: sensingwhether individual seeds are successfully isolated from a bulk of seedsat the seed loading station; and sensing whether the isolated seeds areproperly positioned by the seed transport subsystem adjacent the seedsampling station in preparation for removing tissue from the isolatedseeds.
 2. The method of claim 1, further comprising sensing whether theisolated seeds are properly positioned by the seed transport subsystemadjacent a seed imaging station of the seed sampling station inpreparation for collecting image data of the isolated seeds.
 3. Themethod of claim 2, further comprising, in response to the isolated seedsbeing properly positioned adjacent the seed imaging station, collectingimage data of the isolated seeds at the seed imaging station.
 4. Themethod of claim 3, further comprising sensing whether the isolated seedsare properly positioned by the seed transport subsystem adjacent a seedorientation station of the seed sampling station in preparation fororienting the isolated seeds in a desired orientation.
 5. The method ofclaim 4, further comprising, in response to the isolated seeds beingproperly positioned adjacent the seed orientation station, orienting theisolated seeds at the seed orientation station in the desiredorientation based on the collected image data.
 6. The method of claim 5,further comprising sensing whether the isolated seeds are properlyoriented in the desired orientation at the seed orientation station. 7.The method of claim 5, further comprising removing tissue from theoriented seeds at the seed sampling station.
 8. The method of claim 7,further comprising: receiving the tissue removed from the oriented seedsin a sample tray and mapping a location of the received tissue in thesample tray; and/or receiving the seeds from which the tissue is removedin a seed tray and mapping a location of the seeds in the seed tray. 9.The method of claim 7, further comprising screening the tissue removedfrom the oriented seeds for presence or absence of at least onecharacteristic.
 10. The method of claim 9, further comprising selectingdesired ones of the seeds based on presence or absence of the at leastone characteristic.
 11. The method of claim 1, further comprisingsensing whether the isolated seeds are properly positioned by the seedtransport subsystem adjacent a seed orientation station of the seedsampling station in preparation for orienting the isolated seeds in adesired orientation.
 12. The method of claim 11, further comprising, inresponse to the isolated seeds being properly positioned adjacent theseed orientation station, orienting the isolated seeds at the seedorientation station in the desired orientation.
 13. The method of claim12, further comprising removing tissue from the oriented seeds at theseed sampling station.
 14. The method of claim 13, further comprisingcollecting the tissue removed from the oriented seeds and correlatingthe collected tissue to the seeds from which the tissue is removed. 15.The method of claim 14, further comprising screening the tissue removedfrom the seeds for presence or absence of at least one characteristic.16. The method of claim 15, further comprising selecting desired ones ofthe seeds based on presence or absence of the at least onecharacteristic.
 17. The method of claim 11, further comprising sensingwhether the isolated seeds are properly oriented at the seed orientationstation.
 18. The method of claim 17, further comprising, in response tothe isolated seeds being properly positioned adjacent the seedorientation station and in response to the isolated seeds being in animproper orientation, orienting the isolated seeds at the seedorientation station in the desired orientation.
 19. The method of claim1, further comprising isolating the seeds from the bulk of seeds at theseed loading station.
 20. The method of claim 19, further comprisingdirecting each of the isolated seeds to one of multiple seed holders ofthe seed transport subsystem and transporting the isolated seeds, by theseed transport subsystem, to the seed sampling station.